Cloned (Comment) | Organism |
---|---|
wild-type and mutants cloned into pET3a vectors and overexpressed in Escherichia coli BL21(DE3)pLysS cells | Synechococcus elongatus PCC 7942 = FACHB-805 |
Protein Variants | Comment | Organism |
---|---|---|
A385S | is blue after purification as the wild-type. Partial formation of oxidized FAD is evident after 2 days as with the wild-type. Shows enhanced semiquinone stability | Synechococcus elongatus PCC 7942 = FACHB-805 |
G389N | is yellow-green after purification, significantly less semiquinone present. Reaction time of ca. 3 days is required for complete conversion of semiquinone to oxidized FAD. Kinetic stability of the semiquinone is significantly reduced, semiquinone oxidation rates more closely resemble that in cryptochrome-DASH | Synechococcus elongatus PCC 7942 = FACHB-805 |
M353Q | is blue after purification as the wild-type. Partial formation of oxidized FAD is evident after 2 days as with the wild-type. Has little impact on semiquinone stability | Synechococcus elongatus PCC 7942 = FACHB-805 |
W392Y | is yellow-green after purification, significantly less semiquinone present. Reaction time of ca. 2 days is required for complete conversion of semiquinone to oxidized FAD. Kinetic stability of the semiquinone is significantly reduced, semiquinone oxidation rates more closely resemble that in cryptochrome-DASH | Synechococcus elongatus PCC 7942 = FACHB-805 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Synechococcus elongatus PCC 7942 = FACHB-805 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutants purified by centrifugation, sonication and gel filtration | Synechococcus elongatus PCC 7942 = FACHB-805 |
Storage Stability | Organism |
---|---|
-80°C, 10 mM KH2PO4/K2HPO4 buffer, pH 7, 350 mM NaCl, 10% glycerol | Synechococcus elongatus PCC 7942 = FACHB-805 |
Synonyms | Comment | Organism |
---|---|---|
cyclobutane pyrimidine dimer photolyase | - |
Synechococcus elongatus PCC 7942 = FACHB-805 |
photolyase | - |
Synechococcus elongatus PCC 7942 = FACHB-805 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | large kinetic isotope and pH effects on the rate constants for FAD semiquinone oxidation, which reveal that proton transfer is rate-limiting. Photolyase-specific residues, Trp392 and Gly389, independently ensure a high kinetic barrier to semiquinone reactivity in photolyase, possibly through interactions with the adenine moiety of FAD and/or adjusting the polarity of the binding site. These residues have a much greater impact on semiquinone reactivity than the more FAD proximal Met353 or Ser395 | Synechococcus elongatus PCC 7942 = FACHB-805 |